Cardiac Revascularization with Direct Reprogramming Approaches

通过直接重编程方法进行心脏血运重建

基本信息

批准号：
10557918
负责人：
Young-Sup Yoon
金额：
$ 44.91万
依托单位：
EMORY UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-02-12 至 2025-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10557918
关键词：
Adenovirus Vector Adult American Animals Area Biocompatible Materials Biological Biomedical Engineering Blood Vessels Bone Marrow Cardiac Cardiac Myocytes Cardiovascular Diseases Cell Lineage Cell Reprogramming Cell Survival Cell Therapy Cell Transplantation Cells Clinical Coronary Arteriosclerosis Disease Disease model Echocardiography Embryo Encapsulated Endothelial Cells Endothelium Engineering Ethics Family Fibroblasts Functional disorder Gel Generations Genes Genomics Goals Growth Heart Hepatocyte Histologic Human Impairment Injections Insertion Mutation Ischemia Lentivirus Vector Magnetic Resonance Imaging Medical Modeling Molecular Morbidity - disease rate Mus Myocardial Ischemia Neurons Nitric Oxide Operative Surgical Procedures Patients Peptides Play Population Research Risk Rodent Role Somatic Cell Technology Therapeutic Therapeutic Effect Time Tissue Engineering Tissues Variant Vascularization blood vessel development cardiac regeneration cell type clinical application clinical development clinical translation cost delivery vehicle design heart function improved in vivo induced pluripotent stem cell member microCT mortality nano neovascularization next generation novel novel strategies overexpression peptide amphiphiles postnatal human programs regenerative therapy side effect success transcription factor tumor vector

项目摘要

Project Summary Ischemic heart diseases are the leading cause of morbidity and mortality. The underlying problems of these diseases are loss or dysfunction of blood vessels and insufficient new vessel formation. While cell therapy has emerged as a promising option to promote blood vessel growth, no therapy is yet clinically available and there is much room for improvement. The potential of bone marrow (BM)-derived cells turned out to be minimal9, and embryonic or induced pluripotent stem cell (ESC/iPSC)-derived ECs are difficult to maintain, costly to produce, and may cause side effects. To avoid these problems, a new approach, called direct reprogramming or direct conversion has been developed, in which somatic cells are converted into other lineage cells by overexpression of lineage- or cell-type specific transcription factors (TFs). This approach allows simpler and safer target cell generation and has the potential for more convenient clinical translation. We have attempted this direct reprogramming toward ECs using combinations of seven endothelial-related TFs and demonstrated for the first time that ETV2 alone is sufficient to convert human fibroblasts into ECs. However, since we used a lentiviral vector like others, these rECs have restrictions in clinical applicability. The direct EC reprogramming approach for therapy has two options: cell therapy or direct in vivo reprogramming. For clinical application, both require a safer delivery vector to minimize the possibility of genomic integration. Thus, we developed an adenoviral-ETV2 (Ad-ETV2) vector. Another important obstacle for cell therapy is short- term survival of the transplanted cells. To overcome this problem, we have investigated the potential of biomaterial for prolongation of the cell survival and therapeutic effects. We recently showed that peptide amphiphile (PA) nanomatrix gel is very effective, extending survival of human iPSC-derived ECs longer than 10 months and inducing continuous vessel formation in vivo. In this study, first, we will first develop cell-based therapy. We will generate clinically compatible rECs from human fibroblasts and generate an optimal PA construct combining these rECs with various types of PA nanomatrix gel including a newly developed nitric oxide (NO)-releasing PA. We will then determine the vascularization and therapeutic effects of the selected rEC-PA nanomatrix constructs on rodent ischemic heart disease models. Second, we will investigate whether direct delivery of Ad-ETV2 into cardiac ischemia models can directly reprogram fibroblasts into functional endothelial cells and induce vascularization in vivo. We will use genetically modified mice to track the fate of fibroblasts toward ECs in vivo. The goal of this project is to develop clinically applicable cardiac revascularization strategy using a novel direct reprogramming approach combined with tissue engineering technologies. If successful, this study will provide next-generation platforms for broad areas of research and therapy for ischemic heart diseases.

项目摘要缺血性心脏病是发病率和死亡率的主要原因。这些的根本问题疾病是血管的损失或功能障碍，新血管形成不足。虽然细胞疗法有成为促进血管生长的有前途的选择，尚无临床治疗是改进的空间。骨髓（BM）衍生的细胞的潜力被证明是最小9，并且胚胎或诱导的多能干细胞（ESC/IPSC）衍生的EC很难维持，生产成本高昂，并可能引起副作用。为了避免这些问题，一种新方法，称为直接重编程或直接已经开发了转化，其中体细胞通过过表达转化为其他谱系细胞谱系或细胞类型的特异性转录因子（TFS）的。这种方法允许更简单，更安全的目标单元格产生并有可能进行更方便的临床翻译。我们尝试了这个直接使用七个与内皮相关的TF的组合对EC进行重新编程，并证明了第一个仅ETV2就足以将人的成纤维细胞转换为EC。但是，由于我们使用了慢病毒与其他人一样，这些REC在临床适用性方面也有限制。治疗的直接EC重编程方法有两种选择：细胞疗法或直接体内重编程。对于临床应用，两者都需要更安全的递送载体来最大程度地减少基因组整合的可能性。因此，我们开发了腺病毒-ETV2（AD-ETV2）向量。细胞疗法的另一个重要障碍是短暂移植细胞的期限存活。为了克服这个问题，我们研究了生物材料，用于延长细胞存活和治疗作用。我们最近证明了肽 Amphiphile（PA）Nanomatrix凝胶非常有效，扩展了人IPSC衍生的EC的存活超过10 几个月并在体内诱导连续血管形成。在这项研究中，首先，我们将首先开发基于细胞的治疗。我们将从人类成纤维细胞并生成最佳的PA构建体，将这些REC与各种类型的PA结合在一起纳米质凝胶，包括新开发的一氧化氮（NO） - 释放PA。然后，我们将确定选定的Rec-Pa纳米晶体构建体对啮齿动物缺血性心脏的血管化和治疗作用疾病模型。其次，我们将调查是否将AD-ETV2直接传递到心脏缺血模型中可以直接将成纤维细胞重新编程为功能性内皮细胞，并在体内诱导血管化。我们将使用经过转基因的小鼠以跟踪成纤维细胞朝向ECS体内的命运。该项目的目的是开发临床适用的心脏血运重建策略使用新型的直接重编程方法合并使用组织工程技术。如果成功，这项研究将为广泛的下一代平台提供缺血性心脏病的研究和治疗领域。

项目成果

期刊论文数量（5）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

FLT4 as a marker for predicting prognostic risk of refractory acute myeloid leukemia.

DOI：
10.3324/haematol.2022.282472
发表时间：
2023-11-01
期刊：
HAEMATOLOGICA
影响因子：
10.1
作者：
Lee, Ji Yoon;Lee, Sung-Eun;Han, A-Reum;Lee, Jongeun;Yoon, Young-sup;Kim, Hee-Je
通讯作者：
Kim, Hee-Je